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Peptide Microarrays

Volume 570 of the series Methods in Molecular Biology™ pp 309-316

Date:

A Novel Combinatorial Approach to High-Density Peptide Arrays

  • Mario BeyerAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
  • , Ines BlockAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
  • , Kai KönigAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
  • , Alexander NesterovAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
  • , Simon FernandezAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
  • , Thomas FelgenhauerAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
  • , Christopher SchirwitzAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
  • , Klaus LeibeAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
  • , Ralf F. BischoffAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
    • , Frank BreitlingAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center
    • , Volker StadlerAffiliated withDepartment of Chip-Based Peptide Libraries, German Cancer Research Center

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Abstract

Combinatorial synthesis of peptides on solid supports (1), either as spots on cellulose membranes (2) or with split-pool-libraries on polymer beads (3), substantially forwarded research in the field of peptide–protein interactions. Admittedly, these concepts have specific limitations, on one hand the number of synthesizable peptide sequences per area, on the other hand elaborate decoding/encoding strategies, false-positive results and sequence limitations. We recently established a method to produce high-density peptide arrays on microelectronic chips (4). Solid amino acid microparticles were charged by friction and transferred to defined pixel electrodes onto the chip’s surface, where they couple to a functional polymer coating simply upon melting (Fig. 16.1 AD,F). By applying standard Fmoc chemistry according to Merrifield, peptide array densities of up to 40,000 spots per square centimetre were achieved (Fig. 16.1G). The term “Merrifield synthesis” describes the consecutive linear coupling and deprotecting of L-amino acids modified with base-labile fluorenylmethoxy (Fmoc) groups at the N-terminus and different acid-sensitive protecting groups at their side chains. Removing side chain protecting groups takes place only once at the very end of each synthesis and generates the natural peptide sequence thereby.

Key words

Solid-phase peptide synthesis peptide array combinatorial peptide libraries atom transfer radical polymerization (ATRP) amino acid microparticles peptide chips